Hemoglobin-catalyzed fluorometric method for the determination of glutathione

Abstract

A new spectrofluorometric method for the determination of glutathione based on the reaction catalyzed by hemoglobin was reported. The reaction product gave a highly fluorescent intensity with the excitation and emission wavelengths of 320.0 nm and 413.0 nm, respectively. The optimum experimental conditions were investigated. Results showed that low concentration glutathione enhanced the fluorescence intensity significantly. The line ranges were 1.0 × 10^–6–1.0 × 10^–5 mol L^–1 of glutathione and 6.0 × 10^-10 mol L^-1–1.0 × 10^-8 mol L^-1, respectively. The detection limit was calculated to be 1.1 × 10^-11 mol L^-1. The recovery test by the standard addition method gave values in the range of 90.78%-102.20%. This method was used for the determination of glutathione in synthetic and real samples with satisfactory results.